Anion exchange chromatography – Mass spectrometry for monitoring multiple quality attributes of erythropoietin biopharmaceuticals

Abstract Assessment of critical quality attributes the biopharmaceutical erythropoietin (EPO) prior to product release requires use several analytical methods. We developed an MS-compatible anion exchange (AEX) method for monitoring multiple EPO biopharmaceuticals. AEX was performed using a stationary phase with quaternary ammonium functional groups and pH gradient elution. Baseline separation charge variants high-quality MS data were achieved 30 mM formate 5.5 formic acid 2.5 as mobile phases. In single experiment, assessment attributes, such heterogeneity, sialic content number N-acetyllactosamine units, possible while providing additional information on other modifications O-acetylation deamidation. addition, good repeatability robustness relative areas individual glycoforms average Neu5Ac per molecule observed. The results comparable common pharmacopeia standard methods advantage requiring fewer less sample treatment saving time costs.